title: Recombinant pseudorabies virus DNase exhibits a RecBCD-like catalytic function.
abstract: The pseudorabies virus (PRV) DNase gene has previously been mapped within the PRV genome. To characterize further the enzymic properties of PRV DNase, this enzyme was expressed in Escherichia coli with the use of a pET expression vector. The protein was purified to homogeneity and assayed for nuclease activity in vitro. Recombinant PRV DNase exhibited an alkaline pH preference and an absolute requirement for Mg2+ ions that could not be replaced by Ca2+ and Na+ ions. Further studies showed that PRV DNase exhibited endonuclease, 5'-exonuclease and 3'-exonuclease activities in both single-stranded and double-stranded DNA. This activity occurred randomly and no significant base preference was demonstrated. The multiple biochemical activities of PRV DNase are similar to the activities of Neurospora crassa endo-exonuclease and E. coli RecBCD, two additional enzymes that are involved in recombination. Taken together, the similarity of action between N. crassa endo-exonuclease, E. coli RecBCD, and PRV DNase suggests that PRV DNase might have a role in the process of recombination that occurs during PRV infection.
authors.affiliation: Department of Microbiology, China Medical College, Taichung, Taiwan 40421, ROC.
authors.forename: C Y
authors.initials: CY
authors.lastname: Hsiang
authors.forename: T Y
authors.initials: TY
authors.lastname: Ho
authors.forename: C H
authors.initials: CH
authors.lastname: Hsiang
authors.forename: T J
authors.initials: TJ
authors.lastname: Chang
mesh_terms: D002384:Catalysis; D004268:DNA-Binding Proteins / Q000378:metabolism; D003851:Deoxyribonucleases / Q000378:metabolism*; D004720:Endonucleases / Q000737:chemistry; D043211:Exodeoxyribonuclease V; D005090:Exodeoxyribonucleases / Q000378:metabolism*; D005092:Exonucleases / Q000737:chemistry; D011558:Herpesvirus 1, Suid / Q000201:enzymology*; D008970:Molecular Weight; D011994:Recombinant Proteins; D013379:Substrate Specificity; D014764:Viral Proteins / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D003851:Deoxyribonucleases; D004720:Endonucleases; D005090:Exodeoxyribonucleases; D005092:Exonucleases; D043211:Exodeoxyribonuclease V